Molecular Biology Reports最新文献

Background: Methicillin-resistant Staphylococcus aureus (MRSA) is a significant public health concern due to its resistance to multiple antibiotics, primarily mediated by the gene mecA, which encodes penicillin-binding protein 2a (PBP2a), considering the growing global demand for novel antimicrobial strategies.

Materials and methods: A clinical MRSA strain harboring the gene mecA was identified. Antimicrobial susceptibility testing was performed to determine the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of Shirazi thyme (Zataria multiflora) essential oil. The strain's susceptibility to multiple antibiotics was assessed and the synergistic effect of Z. multiflora essential oil and cefoxitin was also evaluated. Gas chromatography-mass spectrometry (GC-MS) was employed to identify the bioactive compounds in the essential oil. Molecular docking studies were performed to evaluate the competitive binding affinity of those compounds to PBP2x.

Results: The MRSA strain exhibited resistance to all tested antibiotics except linezolid. The MIC and MBC values for Z. multiflora essential oil were 3.125 mg.mL^- 1 and 6.25 mg.mL^- 1, respectively, for the reference S. aureus strain (ATCC 25923), and 6.25 mg.mL^- 1 for both MIC and MBC for the clinical MRSA strain. The synergy assay demonstrated an enhanced inhibition zone for cefoxitin in combination with Z. multiflora essential oil, indicating a synergistic interaction. Molecular docking studies revealed strong binding interactions between spathulenol, isospathulenol, and aromadendrene comparable to clinically used β-lactam antibiotics.

Conclusion: The findings highlight the significant antibacterial activity of Z. multiflora essential oil against MRSA and its potential to enhance the efficacy of cefoxitin through synergistic interactions among the inhibitors of natural origin for PBP2x.

Background: Infertility affects 50-80 million individuals globally (WHO), with male factors, particularly abnormal semen parameters, contributing to 50% of cases. Among these, impaired sperm motility (asthenozoospermia, AZ) critically compromises fertilization potential. This study aimed to identify molecular biomarkers for AZ by investigating gene and competing endogenous RNA (ceRNA) networks associated with sperm motility.

Methods: The study population included 100 participants, including 50 AZ and 50 Normozoospermia (NZ, control), according to the semen criteria (WHO 2021). We integrated bioinformatics analyses, including differential gene expression profiling (Gene Expression Omnibus dataset), protein-protein interaction (PPI) networks, and miRNA-lncRNA-mRNA regulatory networks. Candidate biomarkers were experimentally validated via qRT-PCR and receiver operating characteristic (ROC) curve analyses in AZ patients.

Results: An integrated bioinformatics and experimental approach identified AKT1, hsa-miR-1286, and LINC00242 as a putative network associated with sperm motility that warrants further functional validation. QRT-PCR validation demonstrated significant downregulation of AKT1 and hsa-miR-1286, alongside upregulation of LINC00242 in AZ samples. Expression levels of AKT1 and hsa-miR-1286 showed positive correlations with sperm motility, whereas LINC00242 exhibited a strong negative correlation. ROC curve analysis further suggests preliminary diagnostic potential of these molecules.

Conclusions: Key findings suggested that AKT1, hsa-miR-1286, and linc00242 may represent preliminary diagnostic molecules, with the hsa-miR-1286/linc00242/AKT1 relationship emerging as a putative expression associated network linked to sperm motility.

Background: Cotton (Gossypium hirsutum L.) yield is threatened by heat stress that impair pollen development and reduce reproductive success.

Objectives: This study aimed to identify tolerant genotypes by integrating physiological screening with molecular analysis of key pollen development genes.

Materials and methods: Twenty cotton genotypes were evaluated in a randomized complete block design (RCBD) with three replicates at the research area of MNS-University of Agriculture Multan under normal and heat stress conditions for two consecutive years. Physiological parameters including 2,3,5 triphenyl-tetrazolium chloride (TTC), acetocarmine-based pollen viability and relative cell membrane thermo-stability were measured to assess heat tolerance and sensitive genotypes.

Results: Heat tolerant genotypes including UAM-20, FH-492, MNH-1035 and Cyto-177 exhibited higher pollen viability and reduced cell membrane injury under heat stress. Whereas, FH-901, CIM-448, JS-212 and CRIS-5 A were identified as heat sensitive genotypes due to low pollen viability and higher cell membrane injury. Expression analysis of GhADF7 and GhFLA14 through RT-PCR demonstrated differential expression between heat tolerant and sensitive genotypes.

Discussion: Tolerant genotypes showed higher expression of both genes under heat stress as compared to the sensitive genotypes. These results suggest that integration of physiological screening and gene expression profiling provides a reliable approach to screen heat tolerant genotypes and highlights GhADF7 and GhFLA14 as key factors enabling reproductive thermotolerance.

The escalating prevalence of infertility on a global scale represents a critical public concern, with over 50% cases attributed to male-related factors. Y chromosome microdeletion, within the azoospermia factor region, has emerged as a significant genetic contributor to male infertility, with its occurrence varying from 2% to 24% based on the geographic location and genetic background. The prevalence of such microdeletions in infertile individuals varies broadly, with AZFc being the most frequently reported deletion. The gr/gr partial deletion demonstrates a significant ethnic correlation, highlighting the genetic and demographic factors that may influence microdeletions in the AZF region. Reports also indicate significant geographical variation, with eastern and southern areas exhibiting higher prevalence, in contrast to Europe and Australia. In the Clinical aspect, the deletion is a reliable diagnostic tool for identifying infertility and its genetic causes, making it essential for accurate diagnosis and genetic counselling. Moreover, this review addresses challenges associated with the complex Y-Short Tandem Repeat profile interpretation, as well as their potential implication in forensic and medical genetics. The key focus is to investigate the uniparental inheritance of the Y chromosome, its importance in tracing the male lineage, identification of perpetrators in sexual assault cases, and in investigative genetic genealogy. However, the deletion results in an atypical profile pattern, thereby necessitating careful validation strategies to prevent interpretational errors in forensic and kinship analysis. In conclusion, establishing a standardised screening protocol, a unified testing threshold, and centralising the genetic database are essential for improving health management and forensic justice.

Background: Endometriosis is a chronic, estrogen-dependent inflammatory disease characterized by the presence of endometrial-like tissue outside the uterine cavity and is associated with pain, infertility, and impaired quality of life. Although its etiology is multifactorial, the molecular mechanisms underlying lesion establishment and persistence remain incompletely understood.

Methods: This study investigated differential gene expression in ectopic endometrial tissues obtained from women with surgically and histologically confirmed endometriosis, compared with eutopic endometrium from healthy fertile controls. Targeted quantitative real-time PCR was performed to evaluate genes involved in epigenetic regulation, inflammation, angiogenesis, extracellular matrix remodeling, developmental pathways, and hormone signaling. Bioinformatics analyses, including Gene Ontology (GO) enrichment and protein-protein interaction (PPI) network analysis, were applied to elucidate functional pathways and gene interaction networks. Clinical and biochemical parameters were correlated with disease severity classified according to the revised American Society for Reproductive Medicine (rASRM) staging system.

Results: A total of 35 differentially expressed genes were identified, comprising 18 upregulated and 17 downregulated genes. Upregulated genes were predominantly associated with DNA methylation, inflammatory signaling, and angiogenesis, including DNMT1, DNMT3A, VEGFA, IL6, TNF-α, and MMP9, whereas downregulated genes included developmental and hormone-responsive regulators such as HOXA10, HOXA11, ESR1, PGR, and FOXO1. GO analysis revealed significant enrichment of epigenetic modification, immune response, and extracellular matrix organization among upregulated genes, while downregulated genes were enriched in developmental and transcriptional regulatory processes. PPI network analysis identified key hub genes within interconnected functional modules. Clinical and biochemical parameters, including serum CA-125 and CRP, increased progressively with rASRM stage.

Conclusion: This study highlights coordinated dysregulation of epigenetic, inflammatory, angiogenic, and hormonal pathways in ectopic endometrial tissues, contributing to the molecular pathogenesis of endometriosis. While these findings provide insight into disease-associated molecular networks, further validation in larger and independent cohorts is required to clarify their biological and clinical relevance.

Background: Oxidative stress, apoptosis, and inflammation are interconnected pathological processes. Bryophytes, such as Dicranum scoparium, have evolved unique secondary metabolic systems, including potent antioxidants, to survive extreme environmental stressors such as dedication and UV radiation. This study aimed to characterize the chemical composition of D. scoparium aqueous extract (DSAE) and evaluate its multi-targeted therapeutic potential using an integrated metabolomics and in vitro co-culture approach.

Methods: The chemical fingerprint of DSAE was established via GC-MS using a DB-5MS column (split ratio 1:10). Cytoprotective effects were evaluated in NIH/3T3 fibroblasts across a 100-fold serial concentration range (0.027 ng/mL, 2.7 ng/mL, 270 ng/mL, 2.7 µg/mL, and 27 µg/mL). A fibroblast-macrophage co-culture model was employed to monitor intercellular signaling and the regulation of the NOX2/SOD2 and NF-κB/inflammasome pathways under H₂O₂-induced stress.

Results: GC-MS identified 34 metabolites, including erythritol and GABA-related derivatives. DSAE (0.027 ng/mL) significantly restored cell viability (88.4 ± 4.1% vs. 50.2 ± 3.4% in H₂O₂) and reduced ROS mean fluorescence intensity by 44.6 ± 5.2%. DSAE significantly upregulated HO-1 (1.25 ± 0.14-fold, p < 0.01) and GPx1 (0.88 ± 0.09-fold, p < 0.05). In the co-culture system, DSAE accelerated wound closure speed (3.44 ± 0.26 μm/h) and suppressed the transcription of NLRP3 (1.84 ± 0.21-fold) and IL-6 (1.92 ± 0.25-fold) via NF-κB inhibition.

Conclusions: DSAE exerts multi-targeted cytoprotective effects by modulating the NOX2/SOD2 axis and suppressing intercellular inflammatory crosstalk. These findings suggest that D. scoparium is a valuable source of bioactive metabolites for treating chronic inflammatory and oxidative disorders.

Background: Hepatocellular carcinoma (HCC) remains a major global health burden, partly due to the lack of physiologically relevant in vitro models that accurately recapitulate early host-virus interactions and immune responses. Human Hepatocyte Line 5 (HHL-5) is an immortalized hepatocyte cell line that retains key liver-specific functions. This study aimed to characterize the phenotypic, genetic, and metabolic features of HHL-5 cells and evaluate their suitability as a non-cancerous hepatic model, in comparison with the HCC cell line HepG2.

Methods and results: Morphological and phenotypic assessment of cells showed smaller cell and nuclear areas and slower proliferation with markedly longer doubling time of HHL-5 cells than HepG2 cells. Genomic analyses using whole-exome sequencing revealed enrichment of immune-related pathways in HHL-5 cells, including antigen processing and presentation, whereas HepG2 cells showed predominance of DNA replication pathways. Metabolomic profiling of cells by nuclear magnetic resonance spectroscopy showed hepatocyte-like oxidative profiles of HHL-5 cells, in contrast to the glycolytic phenotypes of HepG2 cells. Moreover, Western blotting for selected proteins showed reduced expression of oncogenic and stress-response markers, including c-Myc, pSTAT3, pNrf2, and select cytochrome P450 enzymes.

Conclusion: Our findings support HHL-5 cells as a robust non-cancerous in vitro model for investigating liver diseases, viral infection, and early events in hepatocarcinogenesis.

Clinical trial registration: Not applicable.